Tau Protein Induces Aberrant Alternative Splicing Changes in PS19 Transgenic Mice

TIAN Xuewen, CHEN Chan, WANG Xiong

Abstract

To explore through big data analysis whether aberrant alternative splicing (AS) events precede tauP301S-induced neurodegenerative phenotype in 6-month-old PS19 mice.   Methods  The original sequencing files of the GSE182170 dataset was downloaded from the European Nucleotide Archive (ENA) database with axel, aligned to the reference genome of the ENSEMBL database by using STAR software, and common AS event analysis and visualization were performed with rMATS and rmats2sashimiplot R packages. RSEM software was utilized for gene transcript quantification, Deseq2, edgeR, and limma R packages were used for differential expression analysis, and clusterProfiler R package was applied for GO enrichment analysis. String and Cytoscape were used for protein-protein interaction (PPI) analysis. Gene expression correlation analysis was performed with ggcorrplot R package. AS events were validated using PCR followed by agarose electrophoresis.   Results  A total of 8 079 AS events were identified with rMATS and 117 significant AS events (ΔPSI>0.1, sequencing coverage >1) were selected eventually. The most frequent type of AS event was skipped exon (SE) (50.43%), followed by alternative 3' splice site (A3SS) and mutually exclusive exons (MXE). GO enrichment analysis revealed that synapse organization genes were aberrantly spliced in SE events and spliceosome genes were spliced in A3SS events. PPI and correlation analyses showed that the splicing factor Snrpn was significantly associated with the largest number of transcripts. Agarose electrophoresis confirmed the aberrant AS event of the Lrp8 gene in PS19 mice.   Conclusion  Dysregulated splicing factors may contribute to tauP301S-induced aberrant AS changes. The study also increases the understanding of the cycling of tau protein and splicing factors in tauopathies.


Keywords: Tauopathy, P301S, Alternative splicing, Exon skipping, Synapse


 

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References


BARALLE F E， GIUDICE J. Alternative splicing as a regulator of development and tissue identity. Nat Rev Mol Cell Biol，2017，18: 437–451. doi: 10.1038/nrm.2017.27.

COOPER T A， WAN L， DREYFUSS G. RNA and disease. Cell，2009， 136: 777–793. doi: 10.1016/j.cell.2009.02.011.

TOLLERVEY J R， WANG Z， HORTOBAGYI T， et al. Analysis of alternative splicing associated with aging and neurodegeneration in the human brain. Genome Res，2011，21: 1572–1582. doi: 10.1101/gr.122226. 111.

HAN S， NA Y， KOH I， et al. Alternative splicing regulation of low-frequency genetic variants in exon 2 of TREM2 in Alzheimer's disease by splicing-based aggregation. Int J Mol Sci，2021，22(18): 9865. doi: 10.3390/ijms22189865.

TYSOE C， WHITTAKER J， XUEREB J， et al. A presenilin-1 truncating mutation is present in two cases with autopsy-confirmed early-onset Alzheimer disease. Am J Hum Genet，1998，62: 70–76. doi: 10.1086/ 301672.

WEGMANN S， BIERNAT J， MANDELKOW E. A current view on Tau protein phosphorylation in Alzheimer's disease. Curr Opin Neurobiol， 2021，69: 131–138. doi: 10.1016/j.conb.2021.03.003.

STRANG K H， CROFT C L， SORRENTINO Z A， et al. Distinct differences in prion-like seeding and aggregation between Tau protein variants provide mechanistic insights into tauopathies. J Biol Chem， 2018，293: 2408–2421. doi: 10.1074/jbc.M117.815357.

YOSHIYAMA Y， HIGUCHI M， ZHANG B， et al. Synapse loss and microglial activation precede tangles in a P301S tauopathy mouse model. Neuron，2007，53: 337–351. doi: 10.1016/j.neuron.2007.01.010.

APICCO D J， ZHANG C， MAZIUK B， et al. Dysregulation of RNA splicing in tauopathies. Cell Rep，2019，29(13): 4377–4388. doi: 10.1016/j. celrep.2019.11.093.

WANG W， CAO Q， TAN T， et al. Epigenetic treatment of behavioral and physiological deficits in a tauopathy mouse model. Aging Cell，2021，20(10): e13456. doi: 10.1111/acel.13456.

SVEEN A， AGESEN T H， NESBAKKEN A， et al. Transcriptome instability in colorectal cancer identified by exon microarray analyses: associations with splicing factor expression levels and patient survival. Genome Med，2011，3: 32. doi: 10.1186/gm248.

TROTTER J H， KLEIN M， JINWAl U K， et al. ApoER2 function in the establishment and maintenance of retinal synaptic connectivity. J Neurosci，2011，31(40): 14413–14423. doi: 10.1523/JNEUROSCI.3135-11. 2011.

WASSER C R， MASIULIS I， DURAKOGLUGIL M S， et al. Differential splicing and glycosylation of Apoer2 alters synaptic plasticity and fear learning. Sci Signal，2014，7(353): ra113. doi: 10.1126/scisignal.2005438.

MAPHIS N M， JIANG S， BINDER J， et al. Whole genome expression analysis in a mouse model of tauopathy identifies MECP2 as a possible regulator of tau pathology. Front Mol Neurosci，2017，10: 69. doi: 10. 3389/fnmol.2017.00069.

DHAENENS C M， TRAN H， FRANDEMICHE M L， et al. Mis-splicing of Tau exon 10 in myotonic dystrophy type 1 is reproduced by overexpression of CELF2 but not by MBNL1 silencing. Biochim Biophys Acta，2011，1812: 732–742. doi: 10.1016/j.bbadis.2011.03.010.

SHULMAN J M， IMBOYWA S， GIAGTZOGLOU N， et al. Functionalscreening in Drosophila identifies Alzheimer's disease susceptibility genes and implicates Tau-mediated mechanisms. Hum Mol Genet，2014，23: 870–877. doi: 10.1093/hmg/ddt478.


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